IEF

On 12 Jan 1998, Robert Woodward wrote:
> Dear All,
>> Does anyone in the Cambridge (UK) area have any experience with IEF?
> I would like to know the IEP of a pertially purified protein that I
> have antibodies to. I only have standard 1D miniprotean II equipment
> and I first of all need to know if this could be used in some way, and
> secondly can you western blot IEF gels and then probe with antibodies so
> as to identify my protein of interest? If you need specialist equipment
> is anyone prepared to run a couple of samples out for me (or loan me some
> equipment)? obviously I would be prepared to pay for materials used etc..
>> Thanks very much
>> Robert Woodward
>> Email rw200 at cus.cam.ac.uk> Cambridge University
> Dept Obs+Gynae
> Rosie Maternity Hospital
> Cambridge
>>
We've had good luck with the protocol of Robertson et al. Anal. Biochem.
167: 290-294 (1987). IEF gels are very fragile (5% polyacrylamide) and
thus would be difficult (but possible) to immunoblot. We use GelBond PAG
support film (FMC Bioproducts) to provide stability to the gels, but you
cannot transfer the gel to blotting paper when it's attached to the film.
Someone used to make a porous support film that permitted transfer, but I
can't remember the company. Perhaps someone in the newsgroup knows a
name.
Good Luck,
Stephen M. Marek
Department of Plant Pathology
University of California, Davis
One Shields Avenue
Davis, CA 95616
lab: (530)752-8015
fax: (530)752-5674
email: smmarek at ucdavis.edu
Note: New Davis area code '530' effective Nov. 1, 1997